Comparative proteomic profiling of Small Extracellular vesicles derived from iPSCs and tissue specific mesenchymal stem cells.

BACKGROUND: Small Extracellular vesicles (EV) are emerging as crucial intercellular messengers that contribute to the physiological processes. EVs contain numerous functional proteins and nucleic acids derived from their parent cells and have different roles depending on their origin. Functionally, EVs transfer these biological materials from the parent cell to the recipient and thus exhibits a novel therapeutic platform for delivering therapeutics molecules to the target tissue. In this regard, EVs derived from stem cells such as Mesenchymal Stem Cells and iPSCs have demonstrated a higher ability to benefit regenerative medicine. Even though these stem cells share some common properties, due to the differences in their origin (cell sources, the hierarchy of potency, etc) the EVs cargo profiling and functionality may vary. METHOD: We used iTRAQ-based proteomic analysis to conduct a comprehensive and quantitative evaluation of EVs derived from iPSCs and various tissue-specific MSCs in this study. Additionally, the data was analyzed using a variety of bioinformatic tools, including ProteinPilot for peptide and protein identification and quantification; Funrich, GO, Reactome, and KEGG (Kyoto Encyclopedia of Genes and Genomes) for pathway enrichment; the STRING database, and the inBio Discover tool for identifying known and predicted Protein-Protein networks. RESULTS: Bioinformatics analysis revealed 223 differentially expressed proteins in these EVs; however, Wharton's jelly MSC-EV contained more exclusive proteins with higher protein expression levels. Additionally, 113 proteins were abundant in MSC-EVs, while others were shared between MSC-EVs and iPSC-EVs. Further, based on an in-depth examination of the proteins, their associated pathways, and their interactions with other proteins, it was determined that these proteins are involved in bone regeneration (9.3%), wound healing (4.4%), immune regulation (8.9%), cardiac regeneration (6.6%), neuro regeneration (8.9%), and hepatic regeneration (3.5%). CONCLUSION: Overall, the results of our proteomic analysis indicate that EVs derived from MSCs have a more robust profile of proteins with higher expression levels than iPSCs. This is a significant finding, as it demonstrates the critical therapeutic role of EVs in a variety of diseases, as demonstrated by enrichment analysis, their versatility, and broad application potential.

Hypoxia preconditioning elicit differential response in tissue-specific MSCs via immunomodulation and exosomal secretion.

Mesenchymal stromal cells (MSCs) are emerging as an ideal candidate for regenerative medicine. It is known that the culture conditions impact the cellular properties of MSCs and their therapeutic behavior. Moreover, maintenance of MSCs in low oxygen tension for a short duration has shown to be beneficial for MSCs as it is similar to that of their physiological niche. However, the precise mechanism through which hypoxia pre-conditioning affects MSCs is not clear yet. Thus, in this study, we have investigated the effect of hypoxia exposure (1% O2) on tissue-specific MSCs over a period of time under serum-free culture conditions and evaluated the changes in expression of immuno-modulatory molecules and exosome biogenesis and secretion markers. It was observed that all MSCs responded differentially towards hypoxia exposure as indicated by the expression of HIF-1α. Moreover, this short-term exposure did not induce any changes in MSCs cellular morphology, proliferation rate, and surface marker profiling. In addition, we observed an enhancement in the expression of immunomodulatory factors (HLA-G, PGE-2, and IDO) after hypoxia exposure of 12 to 24 h in all tissue-specific MSCs. Interestingly, we have also observed the upregulation in exosome secretion that was further corelated to the upregulation of expression of exosome biogenesis and secretion markers (ALIX, TSG101, RAB27a, RAB27b). Though there was a differential response of MSCs where WJ-MSCs and BM-MSCs showed upregulation of these markers at 6-12 h of hypoxia pre-conditioning, while AD-MSCs showed similar changes beyond 24 h of hypoxia exposure.

Engineering Cellular Membrane for Dual Mode Therapy Using NIR Responsive Photosensitizer and Reversible Topoisomerase Inhibition Activity.

Extensive research over past few decades has highlighted the challenges of chemotherapy and prompted the need for multimodality therapy because chemotherapy alone cannot fully eradicate the tumor due to physiological barriers in its effective delivery and systemic side effects. It can be mitigated by adopting nanoparticles as more effective delivery method, but none of them completely prevents drug toxicities. Utilizing multiple therapeutic modes such as phototherapy that can act synergistically with chemotherapy in controlling tumor growth, while reducing the overall dosage, could become a preferred route for cancer management. Careful selection of nanoparticle system, which can simultaneously deliver both drug and photosensitizer, can significantly enhance the therapeutic outcome. Therefore, in this paper, we report development and potential of immune-compatible and long circulating nanoerythrosomes for enhancing the therapeutic potential of camptothecin and indocyanine green against murine cancer model. The RBCs membrane simultaneously loaded the nonpolar drug and amphiphilic photosensitizer in its lipid bilayer, which self-assembled to form stable nanoparticles. These nano constructs absorbed light in the near-infrared region and hence are suitable for targeting deep seated tissues. The dual chemo-phototherapy had great effect on cell viability and had therapeutic value.

Comparative Evaluation of Anti-Fibrotic Effect of Tissue Specific Mesenchymal Stem Cells Derived Extracellular Vesicles for the Amelioration of CCl4 Induced Chronic Liver Injury.

Mesenchymal Stem Cells (MSCs) derived Extracellular Vesicles (EVs) have emerged as an effective candidate for amelioration of liver fibrosis. However, the effect and the mechanisms of MSC-EVs in liver repair remains elusive. In this study, we have evaluated the differential regenerative efficacy of EVs derived from two different human tissue-specific MSCs (Adipose tissue; AD-MSC and Wharton's Jelly; WJ-MSC), in a murine model of chronic liver fibrosis. Mouse model of chronic liver injury was induced by carbon tetrachloride (CCl4) injection, followed by administration of EVs via the tail vein. Both quantitative and qualitative assessment was done to evaluate the hepatic regenerative potential of tissue specific MSC-extracellular vesicles. EVs, regardless of their MSC source, were found to be effective in alleviating chronic liver fibrosis, as demonstrated by macroscopic alterations in the liver. According to the findings of the comprehensive study, there were subtle variations in the tissue specific MSCs-EVs mediated approaches. A greater anti-fibrotic impact was demonstrated by AD-MSC derived EVs through extracellular matrix alteration and hepatocyte proliferation. WJ-MSC EVs, on the other hand, have an anti-inflammatory effect, as evidenced by alterations in the expression of pro- and anti-inflammatory cytokines. Furthermore, cargo profiling of these EVs revealed differences in the miRNA and protein expression, as well as the pathways that they were associated. Comparative overview of regression of fibrosis using tissue specific MSC derived EVs (credits BioRender.com ).